Project/Area Number |
08670006
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Research Category |
Grant-in-Aid for Scientific Research (C)
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Allocation Type | Single-year Grants |
Section | 一般 |
Research Field |
General anatomy (including Histology/Embryology)
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Research Institution | CHIBA UNIVERSITY,SCHOOL OF MEDICINE |
Principal Investigator |
TOYOTA Fumie CHIBA UNIVERSITY,SCHOOL OF MEDICINE,DEPARTMENT OF ANATOMY AND DEVELOPMENTAL BIOLOGY,ASSOCIATE PROFESSOR, 医学部, 助教授 (60009751)
|
Co-Investigator(Kenkyū-buntansha) |
MAEKAWA Mamiko CHIBA UNIVERSITY,SCHOOL OF MEDICINE,DEPARTMENT OF ANATOMY AND DEVELOPMENTAL BIOL, 医学部, 助手 (20181571)
|
Project Period (FY) |
1996 – 1998
|
Project Status |
Completed (Fiscal Year 1998)
|
Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1998: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1997: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1996: ¥900,000 (Direct Cost: ¥900,000)
|
Keywords | ZP3 receptor / sp56 / aquaporin 7 / spermatozoa / spermiogenesis / immuno-cytochemistry / laser confocal microscope / labeled surface replica / 水チャンネル / 細胞相互作用 / 電子顕微鏡 / マウス / ZP3受容体 / 免疫細胞化学 |
Research Abstract |
1. Distribution of ZP3 receptor on the mouse spermatozoon : ZP3 is one of the three glycoproteins in the zona pellucida, and significant for the sperm recognition, binding and induction of acrosome reaction. The localization of ZP3 receptor on the spermatozoa was determined by colloidal gold-conjugated ZP3 and surface replica technique. The gold particles were detected on the outer surface of the plasma membrane covering the dorsal edge of the sperm head. 2. Localization(1), molecular image (2), and formation (3) of sp56 : (1) sp56 has specific affinity for ZP3. By immuno- cytochemical and surface replica techniques, we demonstrated that sp56 localized at the same area, where the ZP3 receptor was demonstrated in 1. It suggests that sp56 satisfies conditions as a ZP3 receptor. (2) Molecular image of sp56 was observed by low angle rotary shadowing method. The octamer of sp56, which is the form in vivo, appeared as a four-leafed clover, having a maximum diameter of approximately 50nm. Each leaf is thought to be composed of a sp56 dimer. (3) During spermiogenesis, sp56 was observed in the acrosome. A question, when sp56 appear on the outer surface of the plasma membrane, remained to be answered. 3. Formation and localization of aquaporin 7 (AQP7), a water channel protein, during spermiogenesis in the rat testis : Small size of spermatozoa is indispensable for penetration through the zona pellucida. Present immunohistochemical study demonstrated that AQP7 first appears in round spermatid. The plasma membrane covering the cytoplasmic mass showed strong immunoreactivity in the elongated spermatid. The result suggests that AQP 7 works as a water channel, since it is localized on the membrane covering the condensing and diminishing cytoplasm.
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